SigmaSpin™ Sequencing Reaction Clean-Up
SIGMA/S5059 - post-reaction clean-up columns
Synonym: sequencing clean up columns
Product Type: Chemical
Comparison of Sequencing Reaction Clean-Up Methods
Single stranded M13MP18 plasmid was sequenced with a -21M13 forward sequencing primer using ABI BigDye® Terminator chemistry. Sequencing reactions were resolved on an ABI Prism® 377 XL instrument with a 48 cm gel cassette containing AutoPAGE™Plus 4.5% acrylamide at 2.88 kV for 7 hrs.
A. Sequencing reactions were precipitated with 70% ethanol and placed on ice for 30 minutes. DNA pellets were dried and resuspended in TE solution prior to electrophoresis.
B. Sequencing reactions were subjected to post-reaction clean-up with SigmaSpin Post-Reaction Clean-Up 96-Well Plates, according to recommended protocol.
Single stranded M13MP18 plasmid was sequenced with a -21M13 forward sequencing primer using ABI BigDye® Terminator chemistry. Sequencing reactions were resolved on an ABI Prism® 377 XL instrument with a 48 cm gel cassette containing AutoPAGE™Plus 4.5% acrylamide at 2.88 kV for 7 hrs.
A. Sequencing reactions were precipitated with 70% ethanol and placed on ice for 30 minutes. DNA pellets were dried and resuspended in TE solution prior to electrophoresis.
B. Sequencing reactions were subjected to post-reaction clean-up with SigmaSpin Post-Reaction Clean-Up 96-Well Plates, according to recommended protocol.
SigmaSpin™ Post-reaction Clean-Up technology comes in two convenient forms, 96-well plates and single spin columns. Each format comes ready for immediate use.
Sequencing reaction was purified using a SigmaSpin 96-well plate.
pFLAG-MAC™ plasmid (Cat. No. E5644) was sequenced using BigDye® Terminator v3.0 chemistry (20 μL total reaction volume; BigDye® premix diluted 1:1 with SeqSaver™ [Cat. No. S3938]). Data was generated on an ABI Prism® 3700 DNA Analyzer with POP-6 polymer and 10× CE Buffer (Cat. No. B4930). PHRED g20 >600.
pFLAG-MAC™ plasmid (Cat. No. E5644) was sequenced using BigDye® Terminator v3.0 chemistry (20 μL total reaction volume; BigDye® premix diluted 1:1 with SeqSaver™ [Cat. No. S3938]). Data was generated on an ABI Prism® 3700 DNA Analyzer with POP-6 polymer and 10× CE Buffer (Cat. No. B4930). PHRED g20 >600.
Sequencing reaction was purified using a SigmaSpin 96-well plate. pFLAG™ MAC plasmid (Cat. No. E5644) was sequenced using BigDye™ Terminator v 3.0 chemistry (20 μl total reaction volume; BigDye premix diluted 1:1 with SeqSaver™ [Cat. No. S3938]). Data was generated on an ABI Prism® 3700 DNA Analyzer with POP-6™ polymer and 10X CE Buffer (Cat. No. B4930). PHRED q20 >600.
| packaging | pkg of 70 ea |
| Quality Level | 400  |
| storage temp. | 2-8°C |
| working volume | 100 μL |
| Application: | Ideal for removing dye-terminator nucleotides and primers from sequencing reactions and radiolabeled nucleotides, primers, and fluorescent dyes from nucleic acid probe labeling reactions. |
| Application: | SigmaSpin™ Sequencing Reaction Clean-Up has been used in the synthesis of antisense RNA probe. |
| Legal Information: | ABI PRISM is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries |
| Legal Information: | SigmaSpin is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | For additional information, please see www.sigma-aldrich.com/seq. |
| Other Notes: | SigmaSpin Post-Reaction Clean-Up Columns are ideal for lower throughput applications, such as clean-up of probe labeling reactions or small numbers of sequencing reactions. These columns can accept sample volumes up to 100 μL. Each column comes with a collection tube to collect the DNA during centrifugation. SigmaSpin Post-Reaction Clean-Up columns provide a fast, simple, and highly efficient method for removing unincorporated dyes, excess salts, and other interfering reaction components. Each column is packed with a pre-hydrated size-exclusion resin, equilibrated with molecular biology grade water, and supplied in our unique column design. The column also includes a snap-cap bottom seal and an o-ring containing screw-cap to ensure that the resin remains hydrated. SigmaSpin has been tested in high-throughput genome centers and core facilities with ABI PRISM® 3700, 3100, 310 and 377. Each well can accept sample volumes up to 20 μL. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 41106300 |